Contributions
Abstract: PB2339
Type: Publication Only
Background
Non–Hodgkin’s Lymphoma (NHL) was a group of lymphoid malignancies with unsatisfactory treatment effect in some aggressive subtypes. Emodin was an anthraquinone with potent anti-cancer activities. However, the molecular mechanism of Emodin repressing aggressive NHL remains to be revealed in detail.
Aims
To explore the active mechanisms of Emodin in aggressive NHL using a bioinformatics method-Functional-activity network (FAN) analysis and subsequently to verify those bioinformatics findings by in vitro cell experiments.
Methods
The initial target of Emodin, DPTs were acquired by DrugBank. Protein Interaction Network Analysis Platform (PINA) was employed to predict the DPT-protein interaction. Protein interaction chart and network derived from the integrated data was forecasted using String JAVA consortium. Top 10 pathways in Emodin associated network were validated and those which were associated with NHL were investigated by KEGG website and selected for further analysis. The cBio Cancer Genomics Portal was applied to investigate the connection of Emodin related-genes in NHL studies. Subsequently, effect of E35, a novel derivative of emodin were investigated on the diffuse large B cell lymphoma (DLBCL) cell lines SU-DHL4. The effect of E35 on SU-DHL4 cell vitality was tested by 3-(4,5-dimethylthiazol-2-yl)-2,5 diphenyltetra-zolium bromide (MTT) and Colony formation assay. Cell apoptosis following E35 treatment was assessed by Annexin V 7-AAD and PE staining. Western blot analysis was used to confirm those molecular targets predicted by FAN analysis.
Results
Emodin was queried on Drugbank database and 4 of its primary direct protein targets (DPT) were identified. Then the 489 DPTs-associated proteins were predicted on Protein Interaction Network Analysis Platform (PINA). Emodin-DPTs and DPTs-targeted proteins/genes were categorized as Emodin-related proteins/genes and submitted to String database for expanding the protein network and KEGG pathways analysis. 3 signal pathways, Pathway in cancer, Viral carcinogenesis and Transcriptional misregulation in cancer, were identified as significant association with Emodin treatment in NHL. Advanced integrated analysis exhibited the bridging that functionally connected Emodin action to key target protein TP53 in NHL. Subsequently, treatment of E35 on SU-DHL4 cells suppressed cell proliferation and induced apoptosis in a time- and dose-dependent manner, with an IC50 of 0.87±0.10μΜ at 48h. Annenxin-V PE and 7-AAD staining indicated raised apoptosis phenomenon with the increasing dose of E35. Apoptotic cells without intervention were 7.70%±1.55%, while 5μM and 10μM E35 treatment for 24h brought them up to 23.01±2.70%、50.23±1.90%. E35 declined TP53 protein expression and inhibited PI3K/AKT pathway in a dose-dependent manner.
Conclusion
Bioinformatics tool FAN analysis could predict key target protein TP53 in Emodin intervention in DLBCL. Further experiments confirmed the bioinformatics findings. All of above showed that combined bioinformatics analysis and experiments offered a novel approach for understanding of mechanisms of Emodin and its derivative action in NHL with convenience and integrity.
Session topic: 19. Non-Hodgkin lymphoma Biology & Translational Research
Keyword(s): Non-Hodgkin's lymphoma, P53
Abstract: PB2339
Type: Publication Only
Background
Non–Hodgkin’s Lymphoma (NHL) was a group of lymphoid malignancies with unsatisfactory treatment effect in some aggressive subtypes. Emodin was an anthraquinone with potent anti-cancer activities. However, the molecular mechanism of Emodin repressing aggressive NHL remains to be revealed in detail.
Aims
To explore the active mechanisms of Emodin in aggressive NHL using a bioinformatics method-Functional-activity network (FAN) analysis and subsequently to verify those bioinformatics findings by in vitro cell experiments.
Methods
The initial target of Emodin, DPTs were acquired by DrugBank. Protein Interaction Network Analysis Platform (PINA) was employed to predict the DPT-protein interaction. Protein interaction chart and network derived from the integrated data was forecasted using String JAVA consortium. Top 10 pathways in Emodin associated network were validated and those which were associated with NHL were investigated by KEGG website and selected for further analysis. The cBio Cancer Genomics Portal was applied to investigate the connection of Emodin related-genes in NHL studies. Subsequently, effect of E35, a novel derivative of emodin were investigated on the diffuse large B cell lymphoma (DLBCL) cell lines SU-DHL4. The effect of E35 on SU-DHL4 cell vitality was tested by 3-(4,5-dimethylthiazol-2-yl)-2,5 diphenyltetra-zolium bromide (MTT) and Colony formation assay. Cell apoptosis following E35 treatment was assessed by Annexin V 7-AAD and PE staining. Western blot analysis was used to confirm those molecular targets predicted by FAN analysis.
Results
Emodin was queried on Drugbank database and 4 of its primary direct protein targets (DPT) were identified. Then the 489 DPTs-associated proteins were predicted on Protein Interaction Network Analysis Platform (PINA). Emodin-DPTs and DPTs-targeted proteins/genes were categorized as Emodin-related proteins/genes and submitted to String database for expanding the protein network and KEGG pathways analysis. 3 signal pathways, Pathway in cancer, Viral carcinogenesis and Transcriptional misregulation in cancer, were identified as significant association with Emodin treatment in NHL. Advanced integrated analysis exhibited the bridging that functionally connected Emodin action to key target protein TP53 in NHL. Subsequently, treatment of E35 on SU-DHL4 cells suppressed cell proliferation and induced apoptosis in a time- and dose-dependent manner, with an IC50 of 0.87±0.10μΜ at 48h. Annenxin-V PE and 7-AAD staining indicated raised apoptosis phenomenon with the increasing dose of E35. Apoptotic cells without intervention were 7.70%±1.55%, while 5μM and 10μM E35 treatment for 24h brought them up to 23.01±2.70%、50.23±1.90%. E35 declined TP53 protein expression and inhibited PI3K/AKT pathway in a dose-dependent manner.
Conclusion
Bioinformatics tool FAN analysis could predict key target protein TP53 in Emodin intervention in DLBCL. Further experiments confirmed the bioinformatics findings. All of above showed that combined bioinformatics analysis and experiments offered a novel approach for understanding of mechanisms of Emodin and its derivative action in NHL with convenience and integrity.
Session topic: 19. Non-Hodgkin lymphoma Biology & Translational Research
Keyword(s): Non-Hodgkin's lymphoma, P53